Comparative Proteomic Analysis of the PhoP Regulon in Salmonella enterica Serovar Typhi Versus Typhimurium

Background: S. Typhi, a human-restricted Salmonella enterica serovar, causes a systemic intracellular infection in humans (typhoid fever). In comparison, S. Typhimurium causes gastroenteritis in humans, but causes a systemic typhoidal illness in mice. The PhoP regulon is a well studied two component (PhoP/Q) coordinately regulated network of genes whose expression is required for intracellular survival of S. enterica. Methodology/Principal Findings: Using high performance liquid chromatography mass spectrometry (HPLC-MS/MS), we examined the protein expression profiles of three sequenced S. enterica strains: S. Typhimurium LT2, S. Typhi CT18, and S. Typhi Ty2 in PhoP-inducing and non-inducing conditions in vitro and compared these results to profiles of $phoP^−/Q^−$ mutants derived from S. Typhimurium LT2 and S. Typhi Ty2. Our analysis identified 53 proteins in S. Typhimurium LT2 and 56 proteins in S. Typhi that were regulated in a PhoP-dependent manner. As expected, many proteins identified in S. Typhi demonstrated concordant differential expression with a homologous protein in S. Typhimurium. However, three proteins (HlyE, STY1499, and CdtB) had no homolog in S. Typhimurium. HlyE is a pore-forming toxin. STY1499 encodes a stably expressed protein of unknown function transcribed in the same operon as HlyE. CdtB is a cytolethal distending toxin associated with DNA damage, cell cycle arrest, and cellular distension. Gene expression studies confirmed up-regulation of mRNA of HlyE, STY1499, and CdtB in S. Typhi in PhoP-inducing conditions. Conclusions/Significance: This study is the first protein expression study of the PhoP virulence associated regulon using strains of Salmonella mutant in PhoP, has identified three Typhi-unique proteins (CdtB, HlyE and STY1499) that are not present in the genome of the wide host-range Typhimurium, and includes the first protein expression profiling of a live attenuated bacterial vaccine studied in humans (Ty800)

Mineralogical and chemical evolution of the Ernest Henry Fe oxide-Cu-Au ore system, Cloncurry district, northwest Queensland, Australia

The Ernest Henry Cu–Au deposit was formed within a zoned, post-peak metamorphic hydrothermal system that overprinted metamorphosed dacite, andesite and diorite (ca 1740–1660 Ma). The Ernest Henry hydrothermal system was formed by two cycles of sodic and potassic alteration where biotite–magnetite alteration produced in the first cycle formed ca 1514±24 Ma, whereas paragenetically later Na–Ca veining formed ca 1529 +11/−8 Ma. These new U–Pbtitanite age dates support textural evidence for incursion of hydrothermal fluids after the metamorphic peak, and overlap with earlier estimates for the timing of Cu–Au mineralization (ca 1540–1500 Ma). A distal to proximal potassic alteration zone correlates with a large (up to 1.5 km) K–Fe–Mn–Ba enriched alteration zone that overprints earlier sodic alteration. Mass balance analysis indicates that K–Fe–Mn–Ba alteration—largely produced during pre-ore biotite- and magnetite-rich alteration—is associated with K–Rb–Cl–Ba–Fe–Mn and As enrichment and Na, Ca and Sr depletion. The aforementioned chemical exchange almost precisely counterbalances the mass changes associated with regional Na–Ca alteration. This initial transition from sodic to potassic alteration may have been formed during the evolution of a single fluid that evolved via alkali exchange during progressive fluid-rock interaction. Cu–Au ore, dominated by co-precipitated magnetite, minor specular hematite, and chalcopyrite as breccia matrix, forms a pipe-like body at the core of a proximal alteration zone dominated by K-feldspar alteration. Both the core and K-feldspar alteration overprint Na–Ca alteration and biotite–magnetite (K–Fe) alteration. Ore was associated with the concentration of a diverse range of elements (e.g. Cu, Au, Fe, Mo, U, Sb, W, Sn, Bi, Ag, F, REE, K, S, As, Co, Ba and Ca). Mineralization also involved the deposition of significant barite, K(–Ba)–feldspar, calcite, fluorite and complexly zoned pyrite. The complexly zoned pyrite and variable K–(Ba)–feldspar versus barite associations are interpreted to indicate fluctuating sulphur and/or barium supply. Together with the alteration zonation geochemistry and overprinting criteria, these data are interpreted to indicate that Cu–Au mineralization occurred as a result of fluid mixing during dilation and brecciation, in the location of the most intense initial potassic alteration. A link between early alteration (Na–Ca and K–Fe) and the later K-feldspathization and the Cu–Au ore is possible. However, the ore-related enrichments in particular elements (especially Ba, Mn, As, Mo, Ag, U, Sb and Bi) are so extreme compared with earlier alteration that another fluid, possibly magmatic in origin, contributed the diverse element suite geochemically independently of the earlier stages. Structural focussing of successive stages produced the distinctive alteration zoning, providing a basis both for exploration for similar deposits, and for an understanding of ore genesis